LOCUS       sequence-482777-        2313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid insert contains the aMFx1 noKREAEA C-terminal tag sequence 
            for use in Combinatorial Golden Gate Assembly (BsaI restriction 
            sites)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2313)
  AUTHORS   Cleaver A, Luo R, Smith OB, Murphy L, Schwessinger B, Brock J
  TITLE     High-throughput optimisation of protein secretion in yeast via an 
            engineered biosensor.
  JOURNAL   Trends Biotechnol. 2025 Apr;43(4):838-867. doi: 
            10.1016/j.tibtech.2024.11.010. Epub 2024 Dec 13.
  PUBMED    39674781
REFERENCE   2  (bases 1 to 2313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.tibtech.2024.11.010"; journalName: "Trends Biotechnol"; 
            date: "2025-04"; volume: "43"; issue: "4"; pages: "838-867"
COMMENT     Sequence Label: C-term_aMFx1_noKREAEA (4a)
FEATURES             Location/Qualifiers
     source          1..2313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        97..199
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             200..859
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(266..285)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      880..974
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1107..1695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1596..1615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1848..1870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1867..1883
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1883
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2292..2309)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2292..2308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(2301..2313,1..10))
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgtgactggg aaaaccctgg cgcttgtggt ggatcacagt tattggagac ttccttggcc
       61 gtacgatggg ttttgtttca ggtatcaatg agacgttgat cggcacgtaa gaggttccaa
      121 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
      181 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
      241 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
      301 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
      361 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg
      421 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
      481 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
      541 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
      601 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
      661 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
      721 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
      781 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
      841 gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag
      901 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
      961 ttattggtga gaatccaagc actagtaaca acttatatcg tatgggggga cgatttgaag
     1021 ccccctaccc atttacgctg acttgacggg acggcggctt tgttgaatag agcgtcagac
     1081 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1141 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1201 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1261 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1321 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1381 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1441 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1501 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1561 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1621 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1681 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1741 ccttttgctc acatgttctt tccagcgaca tatgccatac gccgaactga actccaccta
     1801 atgtcttgga gattgtcgga tagcgcggaa taggattgtg ccctctgagc ggataacaat
     1861 ttcacacagg aaacagctat gaccatgaca catgcatcca ccatcgcaga cttatcatca
     1921 ctgagcctcc acctagccta tggtttcgta catgtagcgt tcaggagttg tagcaaatcc
     1981 cacgctgaac gattggggta acgccttagg attatatcct agcctgtcgg aacacaagca
     2041 cctttcgtct caggtctcag ccattaatac attggctgcc ctggctttaa ttgtaaccaa
     2101 tgccaggatt gagaccgatg agacggagac accaagcacg ccgagaattt acgtttgctg
     2161 ctcatttgtc cacgcacccg tattcccctg ggagtaaaga tacatgctag tctacctgat
     2221 gctatagtga gcctctaaca ggtcaggcgg aatggcactt ccttattcca gatgcgtgcg
     2281 gattatgcat gactggccgt cgttttacaa cgt
//
